LOCUS       sequence-197906-        5234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning vector with temperature sensitive origin of replication..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5234)
  AUTHORS   Phillips GJ
  TITLE     New cloning vectors with temperature-sensitive replication.
  JOURNAL   Plasmid. 1999 Jan;41(1):78-81.
  PUBMED    9887310
REFERENCE   2  (bases 1 to 5234)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid."; 
            date: "1999-01"; volume: "41(1)"; pages: "78-81"
COMMENT     Sequence Label: pTSA29
FEATURES             Location/Qualifiers
     source          1..5234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             111..686
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG
                     FPRQALNRGLPLGFRFSALRHLDPKKLD"
     primer_bind     134..154
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        136..154
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    167..274
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     191..207
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     191..207
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(241..257)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(242..258)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(282..301)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(283..301)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(311..328)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(311..327)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(320..342)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      469..924
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(556..575)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     766..787
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        950..1054
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1055..1123,1124..1915)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1273..1292)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2543..2765
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2813..3763
                     /codon_start=1
                     /gene="rep101 encoding an A56V mutation"
                     /product="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /label=Rep101(Ts)
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     primer_bind     5101..5118
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tccaccgcgg
      181 tggcggccgc tctagaacta gtggatcccc cgggctgcag gaattcgata tcaagcttat
      241 cgataccgtc gacctcgagg gggggcccgg tacccaattc gccctatagt gagtcgtatt
      301 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
      361 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
      421 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
      481 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
      541 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
      601 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
      661 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
      721 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
      781 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
      841 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
      901 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
      961 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     1021 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     1081 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     1141 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     1201 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     1261 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     1321 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     1381 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     1441 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     1501 tttgcacaac atgggggatc atgtaactcg ccttgatcgc tgggaaccgg agctgaatga
     1561 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     1621 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     1681 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     1741 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     1801 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     1861 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     1921 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     1981 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     2041 gttccactga gcgtcagatc cccgacagta agacgggtaa gcctgttgat gataccgctg
     2101 ccttactggg tgcattagcc agtctgaatg acctgtcacg ggataatccg aagtggtcag
     2161 actggaaaat cagagggcag gaactgctga acagcaaaaa gtcagatagc accacatagc
     2221 agacccgcca taaaacgccc tgagaagccc gtgacgggct tttcttgtat tatgggtagt
     2281 ttccttgcat gaatccataa aaggcgcctg tagtgccatt tacccccatt cactgccaga
     2341 gccgtgagcg cagcgaactg aatgtcacga aaaagacagc gactcaggtg cctgatggtc
     2401 ggagacaaaa ggaatattca gcgatttgcc cgagcttgcg agggtgctac ttaagccttt
     2461 agggttttaa ggtctgtttt gtagaggagc aaacagcgtt tgcgacatcc ttttgtaata
     2521 ctgcggaact gactaaagta gtgagttata cacagggctg ggatctattc tttttatctt
     2581 tttttattct ttctttattc tataaattat aaccacttga atataaacaa aaaaaacaca
     2641 caaaggtcta gcggaattta cagagggtct agcagaattt acaagttttc cagcaaaggt
     2701 ctagcagaat ttacagatac ccacaactca aaggaaaagg actagtaatt atcattgact
     2761 agcccatctc aattggtata gtgattaaaa tcacctagac caattgagat gtatgtctga
     2821 attagttgtt ttcaaagcaa atgaactagc gattagtcgc tatgacttaa cggagcatga
     2881 aaccaagcta attttatgct gtgtggcact actcaacccc acgattgaaa accctacaag
     2941 gaaagaacgg acggtatcgt tcacttataa ccaatacgtt cagatgatga acatcagtag
     3001 ggaaaatgct tatggtgtat tagctaaagc aaccagagag ctgatgacga gaactgtgga
     3061 aatcaggaat cctttggtta aaggctttga gattttccag tggacaaact atgccaagtt
     3121 ctcaagcgaa aaattagaat tagtttttag tgaagagata ttgccttatc ttttccagtt
     3181 aaaaaaattc ataaaatata atctggaaca tgttaagtct tttgaaaaca aatactctat
     3241 gaggatttat gagtggttat taaaagaact aacacaaaag aaaactcaca aggcaaatat
     3301 agagattagc cttgatgaat ttaagttcat gttaatgctt gaaaataact accatgagtt
     3361 taaaaggctt aaccaatggg ttttgaaacc aataagtaaa gatttaaaca cttacagcaa
     3421 tatgaaattg gtggttgata agcgaggccg cccgactgat acgttgattt tccaagttga
     3481 actagataga caaatggatc tcgtaaccga acttgagaac aaccagataa aaatgaatgg
     3541 tgacaaaata ccaacaacca ttacatcaga ttcctaccta cataacggac taagaaaaac
     3601 actacacgat gctttaactg caaaaattca gctcaccagt tttgaggcaa aatttttgag
     3661 tgacatgcaa agtaagtatg atctcaatgg ttcgttctca tggctcacgc aaaaacaacg
     3721 aaccacacta gagaacatac tggctaaata cggaaggatc tgaggttctt atggctcttg
     3781 tatctatcag tgaagcatca agactaacaa acaaaagtag aacaactgtt caccgttaca
     3841 tatcaaaggg aaaactgtcc atatgcacag atgaaaacgg tgtaaaaaag atagatacat
     3901 cagagctttt acgagttttt ggtgcattta aagctgttca ccatgaacag atcgacaatg
     3961 taacagatga acagcatgta acacctaata gaacaggtga aaccagtaaa acaaagcaac
     4021 tagaacatga aattgaacac ctgagacaac ttgttacagc tcaacagtca cacatagaca
     4081 gcctgaaaca ggcgatgctg cttatcgaat caaagctgcc gacaacacgg gagccagtga
     4141 cgcctcccgt ggggaaaaaa tcatggcaat tctggaagaa atagcgcttt cagccggcaa
     4201 accggctgaa gccggatctg cgattctgat aacaaactag caacaccaga acagcccgtt
     4261 tgcgggcagc aaaacccgta cttttggacg ttccggcggt tttttgtggc gagtggtgtt
     4321 cgggcggtgc gcgcaagatc cattatgtta aacgggcgag tttacatctc aaaaccgccc
     4381 gcttaacacc atcagaaatc ctcagcgcga ttttaagcac caaccccccc ccgtaacacc
     4441 caaatccata ctgaaagtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc
     4501 agatcacgca tcctcccgac aacacagacc attccgtggc aaagcaaaag ttcagaatca
     4561 ccaactggtc cacctacaac aaagctctca tcaaccgtgg ctccctcact ttctggctgg
     4621 atgatgaggc gattcaggcc tggtatgagt cggcaacacc ttcatcacga ggaaggcccc
     4681 agcgctattc tgatctcgcc atcaccaccg ttctggtgat taaacgcgta ttccggctga
     4741 ccctgcgggc tgcgcagggt tttattgatt ccatttttgc cctgatgaac gttccgttgc
     4801 gctgcccgga ttacaccagt gtcagtaagc gggcaaagtc ggttaatgtc agtttcaaaa
     4861 cgtccacccg gggtgaaatc gcacacctgg tgattgattc caccgggctg aaggtctttg
     4921 gtgaaggcga atggaaagtc agaaagcacg gcaaagagcg ccgtcgtatc tggcgaaagt
     4981 tgcatcttgc tgtggggatc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5041 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5101 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     5161 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     5221 gagcgcaacg caat
//
